Pharmaceutical composition comprising antisense-nucleic acid for prevention and/or treatment of neuronal injury, degeneration and cell death and for the treatment of neoplasms

ABSTRACT

A pharmaceutical composition comprising an effective amount of a compound which is capable from preventing and treating neuronal injury, degeneration, cell death and/or neoplasms in which expression of c-jun, c-fos or jun-B plays a causal role which compound being an antisense nucleic acid or effective derivative thereof, said antisense nucleic acid hybridizing with an area of the messenger RNA (mRNA) and/or DNA encoding c-jun, c-fos or jun-B.

[0001] The present invention is related to a pharmaceutical compositionand a diagnostic agent comprising an effective amount of a compoundwhich is capable of preventing and treating neuronal injury, cell deathand/or neoplasms in which expression of c-jun, c-fos or jun-B plays acausal role, particularly, antisense nucleic acid or -oligonucleotideshybridizing with an area of the messenger RNA (mRNA) and/or DNAcomprising the genes for c-jun, c-fos or jun-B; the use of the compoundfor the preparation of a pharmaceutical composition for the treatment ofneoplasms and/or the prevention and/or treatment of neuronal injury anddegeneration related with the expression of c-jun, c-fos or jun-B.

[0002] Schlingensiepen et al. report in Proceedings of the AmericanAssociation for Cancer Research, Vol. 32, p. 303, Abstract No. 1799, 82.Annual Meeting of the American Association for Cancer Research, Houston,USA, 1991 that c-jun and jun-B genes share high sequence homology withthe v-jun gene. They belong to the immediate early gene group. C-juntogether with c-fos constitutes the DNA binding factor AP-1. C-jun andjun-B expression was inhibited in different cell lines usingphosphorothioate oligodeoxynucleotides. C-jun inhibition stronglyreduced 3H-thymidine incorporation in two mammary carcinoma cell lines,in the rat phaeochromocytoma cell line PC-12 and in NH 3T3 mousefibroblasts. The inhibition of c-jun expression and of c-fos expressionhad very similar effects in the same cell lines inhibition of jun-Bexpression drastically increases 3H-thymidine uptake to more than 10fold. 10-jun is meant to have the characteristics of a proto-oncogenebut jun-B appears to be an anti-oncogene with strong anti-proliferativeaction similar to that of p53. The results suggest that jun-B and c-junto be functional antagonists with regard to their effect on cell growth.This investigation was carried out in order to elucidate the function ofrespective genes and proteins. This abstract does not suggest anytherapeutic concept.

[0003] From the Journal of Cellular Biochemistry, Abstract B 977,Keystone Symposia on Molecular & Cellular Biology, 1993, Schlingensiepenet al. report of two homologues of the proto-oncogene c-jun which havebeen identified in mammals. In that abstract it is speculated that jun-Bmay play a role in cell-differentiation. In order to investigatefunctional questions of the jun-B gene antisense phosophorothioateoligo-deoxynucleotides (S-ODN) have been used to specifically inhibitexpression of c-jun and jun-B in neuronally differentiating PC-12 tumorcells in primary neuronal cell cultures from the rat hippocampus.Western blot analysis revealed specific reductions in the respective Junprotein levels by more than 90% after application of 2 μM S-ODN. Inneuronal cell cultures neurite outgrowth was strongly inhibited afterinhibition of jun-B expression but was enhanced after application ofanti c-jun-S-ODN. Even more drastic changes were observed in neuronallydifferentiating PC-12 tumor cells. The results suggest that jun-B playsa crucial role in cell differentiation while c-jun appears to inhibitdifferentiation. A therapeutic concept is also not available from thatdisclosure.

[0004] From Biomedicine & Pharmacotherapy, Abstract 38, from the 5.International Congress on Differentiation Therapy, Schlingensiepen etal. report also about the results published in Journal of CellularBiochemistry.

[0005] In Developmental Genetics 14: 305-312 (1993) Schlingensiepen etal. report about the induction of the jun-B and/or c-jun transcriptionfactors. The induction is part of the immediate early response todiverse stimuli that induce alterations in cellular programs. In orderto determine functional significance of the jun-B and/or c-juntranscription antisense phosphorothioate oligodeoxynucleotides were usedto inhibit the expression of the genes in proliferating and neuronallydifferentiating cells. In cell culture studies it was found thatinhibition of jun-B expression markedly reduced morphologicaldifferentiation. Conversely, inhibition of c-jun proteins synthesisenhanced morphological differentiation of both primary neurons and PC-12tumor cells.

[0006] EP-A-0 305 929 deals with membranes with bound oligonucleotidesand peptides directly bound onto the membrane. The method forsynthesizing oligonucleotides directly bound onto a membrane provides ameans for generating membrane affinity supports. A modified membrane forthe method of direct synthesis is also provided.

[0007] WO 92/15680 deals with a method and compositions for theselective inhibition of gene expression. Disclosed are methods andcompositions for the selective inhibition gene expression through theapplication of antisense RNA technology. Antisense RNA constructs employthe use of antisense intron DNA corresponding to distinct intron regionsof the gene whose expression is targeted for down-regulation. In anexemplary embodiment a human lung cancer cell line (NCI-H460a) with ahomozygous spontaneous K-ras mutation was transfected with a recombinantplasmid that synthesizes a genomic segment of K-ras in antisenseorientation. Translation of the mutated K-ras m RNA was specificallyinhibited, whereas expression of H-ras and N-ras was unchanged. Athree-fold growth inhibition occurred in H460a cells when expression ofthe mutated ras p21 protein was down-regulated by antisense RNA andcells remained viable. The growth of H460a tumors in nu/nu mice wassubstantially reduced by expressed K-ras antisense RNA.

[0008] Dan Mercola in Prospects for Antisense Nucleic Acid Therapy ofCancer and AIDS, pp. 83-114, 1991 deals with the use of antisense fosRNA and, to a lesser extent, antisense jun RNA. Such antisense RNA hascontributed to understanding of the roles of gene products in cell cyleregulation, differentiation and so on. Progess in the application ofantisense RNA and oligonucleotides to these topics and implications fordiagnostic and therapeutic approaches are considered.

[0009] S. van den Berg in Prospects for Antisense Nucleic Acid Therapyand Cancer and AIDS, pp. 63-70, 1991 deals with antisense fosoligodeoxyribonucleotides suppressing the generation of chromosomalaberrations. The fast induction of the expression product FOS nuclearonco-protein by serum treatment of starved cells was used to test thefunctional stability of antisense oligodeoxyribonucleotides. Unmodifiedoligodeoxyribonucleotides lost their blocking effect with a half-life ofabout 2 hours, modification of the backbone by thioesters extended thehalf-life to about 4 hours. The modified oligodeoxyribonucleotides whereused to unravel a decisive role of FOS in a complex physiologic event:The induction of chromosomal aberrations upon overexpression ofoncogenes like ras and mos and upon irradiation of fibroblasts withUV-light.

[0010] Induction of the c-Fos, Jun-B and/or c-Jun transcription factorsis part of the immediate early response to diverse stimuli that inducealterations in cellular programs. C-jun and c-fos are proto-oncogeneswhose expression is required for induction of cell proliferation whilethe function of the Jun-B transcription factor has remained unclear.

[0011] Neuronal cell injury and cell death due e. g. to hypoxia orhypoglycemia may occur in cause of responses of the cell to diversestimuli inducing alterations in cellular programs.

[0012] It is an object of the present invention to provide apharmaceutical composition for the prevention and/or treatment ofneuronal injury and/or cell death. Surprinsingly, the expression of thec-fos and c-jun gene plays a causal role in neuronal cell injury andcell death due e. g. to hypoxia or hypoglycemia.

[0013] Furthermore, surprisingly, expression of the Jun-B protein isrequired for the differentiation of normal and neoplastic cells andinhibition of c-Jun protein expression enhances the differentiation ofsuch cells. Based on that result the present invention provides apharmaceutical compositon for the treatment of neoplasms by enhancingjun-B expression and/or inhibiting c-jun expression.

[0014] A pharmaceutical composition comprising antisense nucleic acidsor effective derivatives thereof which hybridize with an area of themRNAs or DNA comprising the genes for c-jun, c-fos or jun-B are able tosolve the problems addressed above. The antisense nucleic acid is ableto hybridize with regions of the c-jun, jun-B or c-fos mRNAs. It isunderstood by the skilled person that fragments of the antisense nucleicacids and antisense nucleic acids containing these sequences workaccording to the invention so long as production of the c-Jun and/orc-Fos and/or Jun-B proteins is reduced or inhibited.

[0015] According to the invention the antisense-oligonucleotides areobtainable by solid phase synthesis using phosphite triester chemistryby growing the nucleotide chain in 3′-5′ direction in that therespective nucleotide is coupled to the first nucleotide which iscovalently attached to the solid phase comprising the steps of

[0016] cleaving 5′DMT protecting group of the previous nucleotide,

[0017] adding the respective nucleotide for chain propagation,

[0018] modifying the phosphite group subsequently cap unreacted5′-hydroxyl groups and

[0019] cleaving the oligonucleotide from the solid support,

[0020] followed by working up the synthesis product.

[0021] The chemical structures of oligodeoxy-ribonucleotides are givenin FIG. 1 as well as the respective structures of antisenseoligo-ribonucleotides are given in FIG. 2. The oligonucleotide chain isto be understood as a detail out of a longer nucleotide chain.

[0022] In FIG. 1 lit. B means an organic base such as adenine (A),guanine (G), cytosine (C) and thymine (T) which are coupled via N9 (A,G) or N1(D,T) to the desoxyribose. The sequence of the bases is thereverse complement of the genetic target sequence (mRNA-sequence). Themodifications used are

[0023] 1. Oligodeoxy-ribonucleotides where all R¹ are substituted by

[0024] 1.1 R¹=O

[0025] 1.2 R¹=S

[0026] 1.3 R¹=F

[0027] 1.4 R¹=CH₃

[0028] 1.5 R¹=OEt

[0029] 2. Oligodeoxy-ribonucleotides where R¹ is varied at theinternucleotide phosphates within one oligonucleotide

[0030] where

[0031] B=deoxy-ribonucleotide dA, dC, dG or dT depending on genesequence

[0032] p=internucleotide phosphate

[0033] n=an oligodeoxy-ribonucleotide stretch of length 6-20 bases

[0034] 2.1 R^(1a)=S; R^(1b)=O

[0035] 2.2 R^(1a)=CH₃; R^(1b)=O

[0036] 2.3 R^(1a)=S; R^(1b)=CH₃

[0037] 2.4 R^(1a)=CH₃; R^(1b)=S

[0038] 3. Oligodeoxy-ribonucleotides where R¹ is alternated at theinternucleotide phosphates within one oligonucleotide

[0039] where

[0040] B=deoxy-ribonucleotide dA, dC, dG or dT depending on genesequence

[0041] p=internucleotide phosphate

[0042] n=an oligodeoxy-ribodinucleotide stretch of length 4-12dinucleotides

[0043] 3.2 R^(1a)=S; R^(1b)=O

[0044] 3.2 R^(1a)=CH₃; R^(1b)=O

[0045] 3.3 R^(1a)=S; R^(1b)=CH₃

[0046] 4. Any of the compounds 1.1-1.5; 2.1-2.4; 3.1-3.3 coupled at R²with the following compounds which are covalently coupled to increasecellular uptake

[0047] 4.1 cholesterol

[0048] 4.2 poly(L)lysine

[0049] 4.3 transferrin

[0050] 4.4 folic acid

[0051] 5. Any of the compounds 1.1-1.5; 2.1-2.4; 3.1-3.3 coupled at R³with the following compounds which are covalently coupled to increasecellular uptake

[0052] 5.1 cholesterol

[0053] 5.2 poly(L)lysine

[0054] 5.3 transferrin

[0055] 5.4 folic acid

[0056] In the case of the RNA-oligonucleotides (FIG. 2) are the basis(adenine (A), guanine (G), cytosine (C), uracil (U)) coupled via N9(A,G) or N1 (C,U) to the ribose. The sequence of the basis is thereverse complement of the genetic target sequence (mRNA-sequence). Themodifications in the oligo-nucleotide sequence used are as follows

[0057] 6. oligo-ribonucleotides where all R¹ are substituted by

[0058] 6.1 R¹=O

[0059] 6.2 R¹=S

[0060] 6.3 R¹=F

[0061] 6.4 R₁=CH₃

[0062] 6.5 R¹=OEt

[0063] 7. Oligo-ribonucleotides where R¹ is varied at theinter-nucleotide phosphates within one oligonucleotide

[0064] where

[0065] B=ribonucleotide A, C, G or T depending on gene sequence

[0066] p=internucleotide phosphate

[0067] n=an oligo-ribonucleotide stretch of length 4-20 bases

[0068] 7.1 R^(1a)=S; R^(1b)=O

[0069] 7.2 R^(1a)=CH₃; R^(1b)=O

[0070] 7.3 R^(1a)=S; R^(1b)=CH₃

[0071] 7.4 R^(1a)=CH₃; R^(1b)=S

[0072] 8. Oligo-ribonucleotides where R¹ is alternated at theinternucleotide phosphates within one oligonucleotide

[0073] where

[0074] B=ribonucleotide A, C, G or T depending on gene sequence

[0075] p=internucleotide phosphate

[0076] n=an oligo-ribodinucleotide stretch of length 4-12 dinucleotides

[0077] 8.2 R^(1a)=S; R^(1b)=O

[0078] 8.2 R^(1a)=CH₃; R^(1b)=O

[0079] 8.3 R^(1a)=S; R^(1b)=CH₃

[0080] 9. Any of the compounds 6.1-6.5; 7.1-7.4; 8.1-8.3 coupled at R²with the following compounds which are covalently coupled to increasecellular uptake

[0081] 9.1 cholesterol

[0082] 9.2 poly(L)lysine

[0083] 9.3 transferrin

[0084] 10. Any of the compounds 6.1-6.5; 7.1-7.4; 8.1-8.3 coupled at R³the following compounds are covalently coupled to increased cellularuptake

[0085] 10.1 cholesterol

[0086] 10.2 poly(L)lysine

[0087] 10.3 transferrin

[0088] 11. Any of the compounds 6.1-6.5; 7.1-7.4; 8.1-8.3; 9.1-9.3;10.1-10.3 where all R⁴ are substituted by

[0089] 11.1 R⁴=O

[0090] 11.2 R⁴=F

[0091] 11.3 R⁴=CH₃

[0092] In a preferred embodiment the c-jun antisense nucleic acidcomprising the sequences as identified in the sequence listing, Seq. ID.No. 1-55 and 174-177.

[0093] In a preferred embodiment the jun-B antisense nucleic acids iscomprising the sequences as identified in the sequence listing Seq. IDNo. 56-97 and 178, 179.

[0094] In another preferred embodiment the c-fos antisense nucleic acidis comprising the sequences as identified in the sequence listing underSeq. ID No. 98-173 and 180-185.

[0095] It is possible that one single individual sequence as mentionedabove works as an antisense nucleic acid or oligo-nucleotide structureaccording to the invention. However, it is also possible that one strandof nucleotides comprises more than one of the sequences as mentionedabove directly covalently linked or with other nucleotides covalentlylinked inbetween. Preferably, individual oligonucleotides of thesequences as outlined in the sequence listing are addressed.

[0096] The sequence

5′ GTCCCTATAC GAAC 3′

[0097] served as randomized control sequence.

[0098] In a preferred embodiment of these oligo-nucleotides they arephosphorotioate derivatives.

[0099] Modifications of the antisense-oligonucleotides are advantageoussince they are not as fast destroyed by endogeneous factors when appliedas this is valid for naturally occuring nucleotide sequences. However,it is understood by the skilled person that also naturally occuringnucleotides having the disclosed sequence can be used according to theinvention. In a very preferred embodiment the modification is aphosphorothioate modification.

[0100] The synthesis of the oligodeoxy-nucleotide of the invention isdescribed as an example in a greater detail as follows.

[0101] Oligodeoxy-nucleotides were synthesized by stepwise 5′-additionof protected nucleosides using phosphite triester chemistry. Thenucleotide A was introduced as5′dimethoxy-trityl-deoxyadenosine(N-benzoyl)-N,N′-diisopropyl-2-cyano-ethylphosphoramidite (0.1 M); C was introduced by a5′-dimethoxytrityl-deoxycytidine(N⁴-benzoyl)-N,N′-diisopropyl-2-cyanoethylphosphoramidite; G was introduced as5′-dimethoxy-trityl-deoxyguanosine(N⁸-isobutyryl)-N,N′-diisopropyl-2-cyanoethylphosphoramidite and the T was introduced as5′-dimethodytrityl-deoxythymidine-N,N′-diisopropyl-2-cyanoethylphosphoramidite. The nucleosides were preferably applied in 0.1 Mconcentration dissolved in acetonitrile.

[0102] Synthesis was performed on controlled pore glass particles ofapproximately 150 μm diameter (pore diameter 500 Å) to which the most 3′nucleoside is covalently attached via a long-chain alkylanin linker(average loading 30 μmol/g solid support).

[0103] The solid support was loaded into a cylindrical synthesis column,capped on both ends with filters which permit adequate flow of reagentsbut hold back the solid synthesis support. Reagents were delivered andwithdrawn from the synthesis column using positive pressure of inertgas. The nucleotides were added to the growing oligonucleotide chain in3′→5′ direction. Each nucleotide was coupled using one round of thefollowing synthesis cycle:

[0104] Cleave 5′DMT (dimethoxytrityl) protecting group of the previousnucleotide with 3-chloroacetic acid in dichloro-methane followed bywashing the column with anhydrous acetonitrile. Then simultaneously oneof the bases in form of their protected derivative depending on thesequence was added plus tetrazole in acetonitrile. After reaction thereaction mixture has been withdrawn and the phosphite was oxidized witha mixture of sulfur (S₈) in carbon disulfid/pyridine/-triethylamine.After the oxidation reaction the mixture was fos S-ODN-180; Lane 3:anti-c-fos S-ODN-182. 10 μg of total protein were used per lane.

[0105]FIG. 4

[0106] Effects of different phosphorothioate oligodeoxynucleotides onc-Jun and Jun-B protein expression. A: Western blots of NIH 3T3 celllysate probed with an anti-c-jun antibody. B:SK-BR3 cell lysates, probedwith an anti-jun-B antibody.

[0107] Incubation times with oligodeoxynucleotide were: Lanes 1-3:6 h;Lanes 4-6: 24 h. Lanes 1 and 4: randomized control S-ODN; Lanes 2 and 5:anti-jun-B S-ODN-62; Lane 3 and 6: anti-c-jun S-ODN-13. 10 μg of totalprotein were used per lane.

[0108]FIG. 5

[0109] Effects of different phosphorothioate oligodeoxynucleotides onc-Jun, Jun-B and c-Fos protein expression.

[0110] A: Enzyme-linked immunosorbent assay of rat PC-12 cell lysatesincubated with c-jun (rat specific) antisense oligo-deoxynucleotides174, 175, 176, 177.

[0111] B: Enzyme-linked immunosorbent assay of human SK-Br-3 celllysates incubated with c-jun (human-specific) antisenseoligodeoxynucleotides 1, 7, 13, 17, 20, 23, 26, 31, 31, 39, 45, 51 or54.

[0112] C: Enzyme-linked immunosorbent assay of rat PC-12 cell lysatesincubated with jun-B (rat-specific) antisense oligo-deoxynucleotides 178or 179.

[0113] D: Enzyme-linked immunosorbent assay of human SK-Br-3 celllysates incubated with jun-B (human-specific) antisenseoligodeoxynucleotides 57, 62, 64, 69, 80 85, 89, 92, 95 or 97.

[0114] E: Enzyme-linked immunosorbent assay of rat PC-12 cell lysatesincubated with c-fos (rat-specific) antisense oligo-deoxynucleotides180, 181, 182, 183, 184 or 185.

[0115] F: Enzyme-linked immunosorbent assay of human SK-Br-3 celllysates incubated with c-fos (human-specific) antisense oligonucleotides98, 99, 102, 103, 108, 116, 121, 130, 139, 144, 152, 158, 165, 170 or173.

[0116] Phosphorothioate-oligodeoxynucleotides were used at 2 μMconcentration. Control cells were left untreated (white bars) or treatedwith 2 μM of randomized control phosphorothioate oligonucleotides (greybars).

[0117]FIG. 6

[0118] Survival of rat cerebellar neurons following hypoxia.Phosphorothioate oligonucleotides were used at 1 μM concentration.Control cells were not subjected to hypoxia (white bar). Hypoxia controlcells were either not treated with oligonucleotide (black bar, C) ortreated with the same concentration of randomized controlphosphorothioate oligo-deoxynucleotide (grey bar). Error bars correspondto 1 SD.

[0119]FIG. 7

[0120] Enhanced proliferation arrest after suppression of c-Jun proteinsynthesis and lack of proliferation arrest in NGF treated PC-12 cellsafter suppression of Jun-B protein synthesis. PC-12 cell number after 8days of NGF treatment. Bars represent the mean of 4 values. Grey bars: 2μM randomized control S-ODN; White bars: 2 μM anti-c-jun S-ODN-174;Black bars: 2 μM anti-jun-B S-ODN-179. Error bars correspond to 1 SD.

[0121]FIG. 8

[0122] Morphological differentiation of NGF treated PC-12 cells afterinhibition of c-jun or jun-B protein synthesis.

[0123] A: Control cells not treated with phosphorothioateoligo-deoxynucleotides.

[0124] B: Cells incubated with 2 μM anti-jun-B S-ODN-179.

[0125] C: Cells incubated with 2 μM anti-c-jun S-ODN-174.

[0126] The invention is further explained by he following non-limitingexamples.

EXAMPLE 1 Cell Lines and Proliferation Assays

[0127] NIH 3T3 mouse fibroblasts and SK-Br-3 human mammary carcinomacells were grown in RPMI medium (Gibco) supplemented with 100 U/mlpenicillin, 100 μg/ml streptomycin, 5% FCS. PC-12 rat phaeochromocytomacells were grown in Dulbecco's modified Eagle's medium (DMEM mediumSeromed), supplemented with 100 U/ml penicillin, 100 μg/ml streptomycin,5% FCS.

EXAMPLE 2 Western Blot

[0128] Cells were kept under low serum conditions in RPMI/2% FCS for 3days, trypsinized and preincubated in RPMI/5% FCS/2 μM S-ODN for 5 min.3×10⁶ cells were plated into 260 ml culture flasks and grown for thetimes indicated in RPMI/5% FCS/2 μM S-ODN, trypsinized, spun down andlysed by freezing. SDS-polyacrylamide gel electrophoresis, blotting andchemiluminescence detection were performed according to standardtechniques. Blots were probed with a rabbit anti mouse-c-jun antibody(Oncogene Science) or with a rabbit anti human-jun-B antibody (OncogeneScience) or with a rabbit anti-c-fos antibody (Oncogene Science), usinggoat anti-rabbit IgG-alkaline-phosphatase conjugate (BoehringerMannheim) as second antibody and CSPD (Tropix) for chemiluminescentdetection.

EXAMPLE 3 Enzyme-Linked Immunosorbent Assay (ELISA)

[0129] Cell lysates were diluted in 50 mM carbonate buffer at pH 9.0 andimmobilized on immunon II plates (Dynatech Laboratories, Inc.)overnight. Antigen solution was removed and 200 μl/well phosphatebuffered saline (PBS)/1% BSA/0.02% azide were added to blocknon-specific protein binding. Following incubation at room temperaturefor 2 h solution was removed. After washing with PBS plates were airdried for 3 h. Specific antibodies for c-jun, jun-B or c-fos (Oncogene,Santa Cruz, Biotechnology Inc.) were added at 50 μl/well, diluted inblocking buffer. Following 1 h incubation at room temperature sampleswere removed and subsequently wells were washed four times withPBS/0.05% Tween 20. Then 50 μl of secondary antibody-phosphataseconjugate were added and removed after 1 h. Wells were washed withdiethanolamine buffer (10 mM diethanolamine, 0.5 mM MgCl₂, pH 9.5). 1tablet of Sigma 104 phosphatase substrate was dissolved in 5 mldiethanolamine buffer. 50 μl of the substrate solution were added perwell. The reaction was stopped with 50 μl 0.1 M EDTA (pH 7.5) and plateswere read on a microtitration plate reader.

EXAMPLE 4 Neuronal Survival

[0130] Cerebella were removed from the brains of 8 day old rats understerile conditions and were transferred into 0.1% trypsin, 0.1% DNase inphosphate buffered saline/glucose solution for 15 min at 20° C.,followed by 1.5% soybean trypsin inhibitor (Sigma) for 5 min. Cells weredissociated in a mixture of Dulbecco's modified Eagle's medium and Ham'sF-12 medium (50%/50%, v/v; DMEM F-12, Gibco) supplemented with KCl 25mM, penicillin (5 U/ml), gentamycin (5 μg/ml) and 30 mM glucose. Cellswere centrifuged at 300 x g for 3 min, and resuspended in the samemedium, supplemented with 10% fetal calf serum (Gibco). Cells wereplated in 3 cm dishes (0.5 ml per well) coated with poly-L-lysine (10μg/ml, Sigma) to a density of 1×10⁵ cells/well and transferred to anincubator with humidified atmosphere with 95% O₂/5% CO₂. Cytosinearabinoside (40 μM) was added after 24 h to inhibit glial cellproliferation. On day 16 after seeding, cells were exposed to anoxia for16 h by placing them in a hermetic chamber containing a humidifiedatmosphere with 95% N₂/5% CO₂. The chamber was transferred into anincubator at 37° C. Phosphorothioate oligodeoxynucleotides were added at1 μM concentration 8 h before the onset of anoxia. Neuronal cell injurywas determined 26 h later by staining with trypan blue dye exclusion(incubation with 0.4% trypan blue for 5 min).

EXAMPLE 5 Proliferation of PC-12 Cells After Treatment with NGF andDifferent Phosphorothioate Oligodeoxynucleotides.

[0131] PC-12 cells were plated at a density of 2,500 cells/well in DMEM(Seromed) supplemented with 100 U/ml penicillin, 100 μg/ml streptomycin,5% FCS/2 μM S-ODN. 2 μM S-ODNs were added 6 h after plating. 24 h afterplating, cells were incubated with 10 ng/ml of the 2.5 S subfraction ofnerve growth factor (NGF) (Boehringer Mannheim) for 8 days. Cell numberwas determined by using trypan blue dye exclusion (incubation with 0.4%trypan blue for 5 min) and counting of cells in a Neubauer countingchamber.

EXAMPLE 6 PC-12 Tumor Cell Differentiation

[0132] PC-12 cells were plated at a density of 2,500 cells/well(Seromed) into 96 well microtitration plates coated with poly-L-lysine(10 μg/ml, Sigma) in 100 μl of DMEM supplemented with 100 U/mlpenicillin, 100 μg/ml streptomycin, 5% FCS, S-ODNs were added at 2 μMconcentration 2 h after plating. 6 h after plating, cells were incubatedwith 40 ng/ml of the 2.5 S subfraction of nerve growth factor (NGF)(Boehringer Mannheim) for 11 days.

1 185 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 1TCGGACTATA CTGC 14 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 2 CAGTTCGGAC TATACT 16 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 3 AAGCCTAAGA CGCA 14 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 4 GCCCAAGTTC AACA 14 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 5 TGAAAAGTCG CGGT14 18 base pairs nucleic acid unknown unknown DNA (genomic) YES 6GGTTAATTAA GATGCCTC 18 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 7 TCTCTAAGAG CGCA 14 16 base pairs nucleic acid unknownunknown DNA (genomic) YES 8 ACGTGAGGTT AGTTTG 16 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 9 CACGTGAGGT TAGT 14 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 10 CATAGAACAG TCCG14 16 base pairs nucleic acid unknown unknown DNA (genomic) YES 11CAGTCATAGA ACAGTC 16 18 base pairs nucleic acid unknown unknown DNA(genomic) YES 12 CTTTGCAGTC ATAGAACA 18 14 base pairs nucleic acidunknown unknown DNA (genomic) YES 13 TGCAGTCATA GAAC 14 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 14 GGTCGTTTCC ATCT 14 16base pairs nucleic acid unknown unknown DNA (genomic) YES 15 CATAGAAGGTCGTTTC 16 14 base pairs nucleic acid unknown unknown DNA (genomic) YES16 CGTCATAGAA GGTC 14 15 base pairs nucleic acid unknown unknown DNA(genomic) YES 17 CATCGTCATA GAAGG 15 24 base pairs nucleic acid unknownunknown DNA (genomic) YES 18 GGACGGGAGG AACGAGGCGT TGAG 24 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 19 TAGCCATAAG GTCC 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 20 GGTTACTGTAGCCA 14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 21GGTTACTGTA GCCA 14 30 base pairs nucleic acid unknown unknown DNA(genomic) YES 22 CAGGGTCATG CTCTGTTTCA GGATCTTGGG 30 18 base pairsnucleic acid unknown unknown DNA (genomic) YES 23 AGTTCTTGGC GCGGAGGT 1818 base pairs nucleic acid unknown unknown DNA (genomic) YES 24AGGTGAGGAG GTCCGAGT 18 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 25 TGGACTGGAT TATCAG 16 18 base pairs nucleic acid unknownunknown DNA (genomic) YES 26 GTGGTGGTGA TGTGCCCG 18 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 27 TGTCACGTTC TTGG 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 28 CTCATCTGTCACGT 14 18 base pairs nucleic acid unknown unknown DNA (genomic) YES 29CGAAGCCCTC GGCGAACC 18 24 base pairs nucleic acid unknown unknown DNA(genomic) YES 30 GCGTGTTCTG GCTGTGCAGT TCGG 24 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 31 CTGCCCCGTT GACC 14 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 32 AGGTTTGCGT AGAC14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 33GGTTGAAGTT GCTG 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 34 CTGGGTTGAA GTTG 14 24 base pairs nucleic acid unknownunknown DNA (genomic) YES 35 TGCTGGGGTT GCGCGGGAAA GGCC 24 20 base pairsnucleic acid unknown unknown DNA (genomic) YES 36 TGCTGCACGG GCATCTGCTG20 27 base pairs nucleic acid unknown unknown DNA (genomic) YES 37GGCACTGTCT GAGGCTCCTC CTTCAGG 27 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 38 ACTCCATGTC GATG 14 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 39 CTCTCCGCCT TGATCC 16 16 basepairs nucleic acid unknown unknown DNA (genomic) YES 40 GTTCCTCATGCGCTTC 16 14 base pairs nucleic acid unknown unknown DNA (genomic) YES41 CTGAGCTTTC AAGG 14 26 base pairs nucleic acid unknown unknown DNA(genomic) YES 42 GCGATTCTCT CCAGCTTCCT TTTTCG 26 30 base pairs nucleicacid unknown unknown DNA (genomic) YES 43 CTGAGCTTTC AAGGTTTTCACTTTTTCCTC 30 14 base pairs nucleic acid unknown unknown DNA (genomic)YES 44 TCCCTGAGCA TGTT 14 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 45 TCTGTTTAAG CTGTGC 16 18 base pairs nucleic acid unknownunknown DNA (genomic) YES 46 CTTTCTGTTT AAGCTGTG 18 16 base pairsnucleic acid unknown unknown DNA (genomic) YES 47 GGTTCATGAC TTTCTG 1614 base pairs nucleic acid unknown unknown DNA (genomic) YES 48CGTGGTTCAT GACT 14 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 49 ACTGTTAACG TGGTTC 16 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 50 CCACTGTTAA CGTG 14 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 51 CCCACTGTTA ACGT 14 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 52 AGCATGAGTT GGCA14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 53GCGTTAGCAT GAGT 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 54 GTTTGCAACT GCTG 14 18 base pairs nucleic acid unknownunknown DNA (genomic) YES 55 CAAAATGTTT GCAACTGC 18 18 base pairsnucleic acid unknown unknown DNA (genomic) YES 56 TCCATTTTAG TGCACATC 1818 base pairs nucleic acid unknown unknown DNA (genomic) YES 57CTGTTCCATT TTAGTGCA 18 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 58 GTGTATGAGT CGTC 14 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 59 CTGTGTATGA GTCG 14 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 60 CGTAGCTGTG TATG 14 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 61 TCGTGTAGAG AGAG14 18 base pairs nucleic acid unknown unknown DNA (genomic) YES 62AGTTTGTAGT CGTGTAGA 18 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 63 GTTTGTAGTC GTGTAG 16 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 64 AGTTTGTAGT CGTG 14 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 65 GGAGTTTGTA GTCG 14 16 basepairs nucleic acid unknown unknown DNA (genomic) YES 66 TCAGGAGTTTGTAGTC 16 18 base pairs nucleic acid unknown unknown DNA (genomic) YES67 GTTTCAGGAG TTTGTAGT 18 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 68 TCGGTTTCAG GAGT 14 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 69 TTGAGACTCC GGTA 14 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 70 ACCAGAAAAG TAGCTG 16 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 71 CCTGACCAGA AAAG14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 72ATTCAGGCGT TCCA 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 73 CTGTTGGGGA CAAT 14 16 base pairs nucleic acid unknownunknown DNA (genomic) YES 74 GGTAAAAGTA CTGTCC 16 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 75 GGGTAAAAGT ACTGTC 16 18 basepairs nucleic acid unknown unknown DNA (genomic) YES 76 GCACCTCCACCGCTGCCA 18 18 base pairs nucleic acid unknown unknown DNA (genomic) YES77 CTCCTGCTCC TCGGTGAC 18 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 78 GCTTTGACAA AGCC 14 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 79 CTTGTGCAGA TCGT 14 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 80 TCATCTTGTG CAGATC 16 16 basepairs nucleic acid unknown unknown DNA (genomic) YES 81 GTTCATCTTGTGCAGA 16 14 base pairs nucleic acid unknown unknown DNA (genomic) YES82 CGTGGTTCAT CTTG 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 83 TCACGTGGTT CATC 14 18 base pairs nucleic acid unknownunknown DNA (genomic) YES 84 GCCCAGGGAC ACGTTGGG 18 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 85 GGTTGGTGTA AACG 14 20base pairs nucleic acid unknown unknown DNA (genomic) YES 86 TACGAGCTCCCGGTCCCGAC 20 14 base pairs nucleic acid unknown unknown DNA (genomic)YES 87 TAGCTGATGG TGGT 14 30 base pairs nucleic acid unknown unknown DNA(genomic) YES 88 CAGCTGCGCC GGGTGGCCAC CGGCGAAGGG 30 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 89 TCCTTGAAGG TGGA 14 16base pairs nucleic acid unknown unknown DNA (genomic) YES 90 TCTTCCATGTTGATGG 16 14 base pairs nucleic acid unknown unknown DNA (genomic) YES91 CTTTGATGCG CTCT 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 92 CTCCACTTTG ATGC 14 30 base pairs nucleic acid unknownunknown DNA (genomic) YES 93 GCTCCAGCTT CCGCTTCCGG CACTTGGTGG 30 30 basepairs nucleic acid unknown unknown DNA (genomic) YES 94 GGCCTTGAGCGTCTTCACCT TGTCCTCCAG 30 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 95 TGACCTTCTG TTTGAG 16 16 base pairs nucleic acid unknownunknown DNA (genomic) YES 96 CATGACCTTC TGTTTG 16 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 97 GTCATGACCT TCTG 14 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 98 CGAGAACATC ATCG14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 99GTAGTCTGCG TTGA 14 20 base pairs nucleic acid unknown unknown DNA(genomic) YES 100 GCTGCAGCGG GAGGATGACG 20 16 base pairs nucleic acidunknown unknown DNA (genomic) YES 101 AGTAAGAGAG GCTATC 16 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 102 GTAGTAAGAG AGGC 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 103 GGTAGTAAGAGAGG 14 16 base pairs nucleic acid unknown unknown DNA (genomic) YES 104GTGAGTGGTA GTAAGA 16 18 base pairs nucleic acid unknown unknown DNA(genomic) YES 105 GTCCGTGCAG AAGTCCTG 18 16 base pairs nucleic acidunknown unknown DNA (genomic) YES 106 GAATGAAGTT GGCACT 16 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 107 GGAATGAAGT TGGC 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 108 GGGAATGAAGTTGG 14 30 base pairs nucleic acid unknown unknown DNA (genomic) YES 109GCTGCACCAG CCACTGCAGG TCCGGACTGG 30 30 base pairs nucleic acid unknownunknown DNA (genomic) YES 110 CTGGTCTGCG ATGGGGCCAC AGAGGAGACG 30 16base pairs nucleic acid unknown unknown DNA (genomic) YES 111 TCATGGTCTTCACAAC 16 30 base pairs nucleic acid unknown unknown DNA (genomic) YES112 CAATGCTCTG CGCTCGGCCT CCTGTCATGG 30 14 base pairs nucleic acidunknown unknown DNA (genomic) YES 113 CTAGAGTTCC TCAC 14 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 114 GAGTACGCTA GAGT 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 115 GAAGAGTACGCTAG 14 28 base pairs nucleic acid unknown unknown DNA (genomic) YES 116CTGCTTCCCA CCCAGCCCCC ACATTCCC 28 20 base pairs nucleic acid unknownunknown DNA (genomic) YES 117 TTCATCCTCT GTACTGGGCT 20 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 118 GTTACGGATG TGCA 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 119 CAGTTACGGATGTG 14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 120CCAGTTACGG ATGT 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 121 AGAGTCTGAG TTGG 14 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 122 GTGAGACTCA GAGT 14 14 base pairs nucleicacid unknown unknown DNA (genomic) YES 123 TCTTAGGGTG AGAC 14 16 basepairs nucleic acid unknown unknown DNA (genomic) YES 124 GAGAGTACTTCTTAGG 16 18 base pairs nucleic acid unknown unknown DNA (genomic) YES125 GGAAGAAACT ATGAGAGT 18 18 base pairs nucleic acid unknown unknownDNA (genomic) YES 126 CTTAGGGAAG AAACTATG 18 16 base pairs nucleic acidunknown unknown DNA (genomic) YES 127 CGGTAAGAAA CTTAGG 16 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 128 AGCATGCGGT AAGA 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 129 GTCTGAAAGCATGC 14 16 base pairs nucleic acid unknown unknown DNA (genomic) YES 130AGAACAAAGA AGAGCC 16 18 base pairs nucleic acid unknown unknown DNA(genomic) YES 131 CAAGAGAACA AAGAAGAG 18 16 base pairs nucleic acidunknown unknown DNA (genomic) YES 132 CAGCAAGAGA ACAAAG 16 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 133 TCCTCAGCAA GAGA 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 134 AGGTGTGACTTGCA 14 16 base pairs nucleic acid unknown unknown DNA (genomic) YES 135GAATAGGTGT GACTTG 16 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 136 CAGAATAGGT GTGACT 16 14 base pairs nucleic acidunknown unknown DNA (genomic) YES 137 GCAGAATAGG TGTG 14 16 base pairsnucleic acid unknown unknown DNA (genomic) YES 138 CAGTTGCAGA ATAGGT 1616 base pairs nucleic acid unknown unknown DNA (genomic) YES 139GAAACCATTT CTGACC 16 18 base pairs nucleic acid unknown unknown DNA(genomic) YES 140 TGTGAAACCA TTTCTGAC 18 18 base pairs nucleic acidunknown unknown DNA (genomic) YES 141 CACTGTGAAA CCATTTCT 18 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 142 CCACTGTGAA ACCA14 30 base pairs nucleic acid unknown unknown DNA (genomic) YES 143AGAACTGGCT CCTGCAGCTT CCCTGCTTCC 30 15 base pairs nucleic acid unknownunknown DNA (genomic) YES 144 CACCTCCATT CACCC 15 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 145 CAGTAAAAGT GTCTGC 16 18 basepairs nucleic acid unknown unknown DNA (genomic) YES 146 CGACATTCAGTAAAAGTG 18 14 base pairs nucleic acid unknown unknown DNA (genomic) YES147 GACCGACATT CAGT 14 18 base pairs nucleic acid unknown unknown DNA(genomic) YES 148 CTTCTGGAGA TAACTAGA 18 18 base pairs nucleic acidunknown unknown DNA (genomic) YES 149 CATCTTATTC CTTTCCCT 18 16 basepairs nucleic acid unknown unknown DNA (genomic) YES 150 CAGCCATCTTATTCCT 16 16 base pairs nucleic acid unknown unknown DNA (genomic) YES151 TGCAGCCATC TTATTC 16 15 base pairs nucleic acid unknown unknown DNA(genomic) YES 152 GAGTGTATCA GTCAG 15 14 base pairs nucleic acid unknownunknown DNA (genomic) YES 153 GGAGTGTATC AGTC 14 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 154 CTTGGAGTGT ATCAGT 16 14 basepairs nucleic acid unknown unknown DNA (genomic) YES 155 ACAGAGTACC TACC14 16 base pairs nucleic acid unknown unknown DNA (genomic) YES 156CCAACTTTCC CTTAAG 16 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 157 CCTTATGCTC AATCTC 16 14 base pairs nucleic acidunknown unknown DNA (genomic) YES 158 GTCTTACTCA AGGG 14 16 base pairsnucleic acid unknown unknown DNA (genomic) YES 159 ACAGTCTTAC TCAAGG 1618 base pairs nucleic acid unknown unknown DNA (genomic) YES 160CATAAGACAC AGTCTTAC 18 18 base pairs nucleic acid unknown unknown DNA(genomic) YES 161 GAAAGCATAA GACACAGT 18 16 base pairs nucleic acidunknown unknown DNA (genomic) YES 162 GGAAAGCATA AGACAC 16 16 base pairsnucleic acid unknown unknown DNA (genomic) YES 163 AGGGATAAAG GAAAGC 1614 base pairs nucleic acid unknown unknown DNA (genomic) YES 164CCTGTATACA GAGG 14 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 165 TGTCTCCTGT ATACAG 16 16 base pairs nucleic acidunknown unknown DNA (genomic) YES 166 CATCTTCTAG TTGGTC 16 16 base pairsnucleic acid unknown unknown DNA (genomic) YES 167 CTCATCTTCT AGTTGG 1618 base pairs nucleic acid unknown unknown DNA (genomic) YES 168CTTCTCATCT TCTAGTTG 18 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 169 CAAAGCAGAC TTCTCA 16 14 base pairs nucleic acidunknown unknown DNA (genomic) YES 170 CTGCAAAGCA GACT 14 18 base pairsnucleic acid unknown unknown DNA (genomic) YES 171 CTAGTTTTTC CTTCTCCT18 18 base pairs nucleic acid unknown unknown DNA (genomic) YES 172TCTAGTTTTT CCTTCTCC 18 16 base pairs nucleic acid unknown unknown DNA(genomic) YES 173 CAGGATGAAC TCTAGT 16 14 base pairs nucleic acidunknown unknown DNA (genomic) YES 174 TCGTAGAAGG TCGT 14 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 175 AGGGTTACTG TAGC 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 176 GTAGTGGTGATGTG 14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 177CGTCGTAGAA GGTC 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 178 TTTCGTGCAC ATCC 14 18 base pairs nucleic acid unknownunknown DNA (genomic) YES 179 AGTTTGTAGT CGTGAAGA 18 14 base pairsnucleic acid unknown unknown DNA (genomic) YES 180 CGAGAACATC ATGG 14 14base pairs nucleic acid unknown unknown DNA (genomic) YES 181 GTAGTAGGAAAGGC 14 14 base pairs nucleic acid unknown unknown DNA (genomic) YES 182GGTAGTAGGA AAGG 14 14 base pairs nucleic acid unknown unknown DNA(genomic) YES 183 GGAATGGTAG TAGG 14 15 base pairs nucleic acid unknownunknown DNA (genomic) YES 184 GGTCATTGAG AAGAG 15 16 base pairs nucleicacid unknown unknown DNA (genomic) YES 185 GCTAATGTTC TTGACC 16

1. A pharmaceutical composition comprising an effective amount of acompound which is capable from preventing and treating neuronal injury,degeneration, cell death and/or neoplasms in which expression of c-jun,c-fos or jun-B plays a causal role which compound being an antisensenucleic acid or effective derivative thereof, said antisense nucleicacid hybridizing with an area of the messenger RNA (mRNA) and/or DNAcomprising the genes c-jun, c-fos or jun-B.
 2. The pharmaceuticalcomposition of claim 1 wherein the c-jun antisense nucleic acid ornucleotide is comprising the sequences as identified in the sequencelisting under Seq. ID No. 1-55 and 174-177 the jun-B antisense nucleicacid comprising the sequence as identified in sequence listing underSeq. ID No. 56-97 and 178-179, and the c-fos antisense nucleic acidcomprising the sequence as identified in the sequence listing under Seq.ID No. 98-173 and 180-185, wherein any sequence listed above representsa single oligonucleotide or represents a section of linked nucleotidesin an nucleic acid and/or a nucleic acid comprising at least one of theabove listed nucleotide sequences directly covalently linked or withother nucleotides separating the nucleotide sequences listed above. 3.The pharmaceutical composition of claims 1 and/or 2, wherein theoligonucleotides are modified oligo-nucleotides such as phosphorothioatederivatives.
 4. Antisense oligonucleotides of the pharmaceuticalcomposition according to claim 3 and 4 as intermediate products formanufacturing the pharmaceutical composition of claims 2 and/or
 3. 5.Antisense nucleic acid or -oligonucleotides according to any one of theclaims 2 to 4 obtainable by solid phase synthesis using phosphitetriester chemistry by growing the nucleotide chain in 3′-5′ direction inthat the respective nucleotide is coupled to the first nucleotide whichis covalently attached to the solid phase comprising the steps ofcleaving 5′DMT protecting group of the previous nucleotide, adding therespective nucleotide for chain propagation, modifying phosphite groupssubsequently cap unreacted 5′-hydroxyl groups and cleaving theoligonucleotide from the solid support, followed by working up thesynthesis product.
 6. Use of a compound according to any one of theclaims 1 to 5 for the preparation of a pharmaceutical composition forthe treatment of neoplasms and/or the prevention and treatment ofneuronal injury and de-generation related with the expression of c-jun,c-fos or jun-B.
 7. Method of treating of neoplasms and/or preventing andtreating of neuronal injury and/or degeneration by administering aneffective amount of the pharmaceutical composition or the compoundaccording to any one of the claims 1 to 5 to a patient suffering fromdisorders related with the expression of c-jun, c-fos or jun-B.
 8. Adiagnostic agent comprising a compound of the claims 1 to 5.